Anti-RPS9 monoclonal antibody Made to order

Aim: Retinoic acid (RA) has proven to possess modest but distinct activity in metastatic renal cell carcinoma (RCC), at least in a subgroup of patients. However, the exact molecular mechanisms leading to success or failure of RA application in individual patients are still unknown. As earlier studies have indicated that in RCC the RA receptor (RAR) beta might play a central role in RA signaling, we investigated the expression of the isoforms RAR-beta(1+2) in primary conventional and chromophobe RCC. Methods: We used quantitative RT-PCR methodology to study RAR-beta(1) and RAR-beta(2) expression in ten primary conventional RCC samples (clear cell type), in two chromophobe RCC specimens, and the respective corresponding normal kidney tissues. The housekeeping genes RPS9 and RPLP0 were applied to normalize differences in mRNA quality and quantity. Results: In contrast to conventional RCC samples, RAR-beta(1) was significantly overexpressed in both chromophobe tumors compared to the adjacent normal kidney tissue (p = 0.03). On the contrary, RAR-beta(2) expression did neither differ significantly between conventional and chromophobe RCC (p = 0.91) nor between malignant and normal kidney tissue (p >= 0.47). Conclusion: We demonstrate for the first time a significant and specific overexpression of RAR-beta(1) in chromophobe RCC. In future we will have to confirm this result within a larger number of samples.

Quantitative Real-Time PCR (qPCR), a sensitive and commonly used technique for gene expression analysis, requires stably expressed reference genes for normalization of gene expression. Up to now, only one reference gene for qPCR analysis, corresponding to 40S Ribosomal protein S9 (RPS9), was available for the medicinal plant Catharanthus roseus, the only source of the commercial anticancer drugs vinblastine and vincristine. Here, we screened for additional reference genes for this plant species by mining C roseus RNA-Seq data for orthologs of 22 genes known to be stably expressed in Arabidopsis thaliana and qualified as superior reference genes for this model plant species. Based on this, eight candidate C roseus reference genes were identified and, together with RPS9, evaluated by performing qPCR on a series of different C roseus explants and tissue cultures. NormFinder, geNorm and BestKeeper analyses of the resulting qPCR data revealed that the orthologs of At2g28390 (SAND family protein, SAND), At2g32170 (N2227-like family protein, N2227) and At4g26410 (Expressed protein, EXP) had the highest expression stability across the different C roseus samples and are superior as reference genes as compared to the traditionally used RPS9. Analysis of publicly available C roseus RNA-Seq data confirmed the expression stability of SAND and N2227, underscoring their value as reference genes for C roseus qPCR analysis. (C) 2014 Elsevier Masson SAS. All rights reserved.